5486
B. Scheiper et al. / Bioorg. Med. Chem. Lett. 21 (2011) 5480–5486
Table 6
Further profiling of selected Renin inhibitorsa
Cpds Renin
IC50
Renin IC50 with
M] human plasma [
Renin IC50 with
mouse plasma [
Caco-2 Permeability
Papp [⁄10ꢀ7 cm/s]
Log P
(MoKa)
pKa (basic, Log D (pH
Ligand
efficiency
Lipophilic
ligand efficiency
[l
l
M]
lM]
MoKa)
7.4, MoKa)
3l
3m
6f
0.014
0.0013
0.014
0.003
0.030
nd
0.025
0.226
0.015
0.036
0.026
0.192
2.8
43
12.7
39
2.1
2.3
2.5
3.7
8.1
8.1
8.1
8.1
1.3
1.5
1.7
2.9
0.33
0.37
0.33
0.30
5.75
6.59
5.35
4.82
7d
a
Data were obtained as described in text and ref. 22. nd = not determined.
fluorogenic substrate Dabcyl-c-Abu-Ile-His-Pro-Phe-His-Leu-Val-Ile-His-Thr-
EDANS is cleaved at the Leu–Val bond in analogy to angiotensinogen. Human
renin at a concentration of 5 nM is incubated with the test compound at
Acknowledgements
The authors would like to thank Sabrina Berger, Holger Gaul,
Silvia Hein, Laetitia Martin, Andrea Müller, Mathilde Pierre, and
Katja Wittmann for their technical assistance.
various concentrations and the substrate at a concentration of 10 lM (2 h at
RT) in 0.05 M Tris buffer (pH 8). The increase in fluorescence due to
fluorescence resonance energy transfer at an emission wavelength of 485 nm
is recorded in a microplate spectrofluorometer (7).
16. McMartin, C.; Bohacek, R. S. J. Comput. Aided Mol. Des. 1997, 11, 333.
17. Friesner, R. A.; Banks, J. L.; Murphy, R. B.; Halgren, T. A.; Klicic, J. J.; Mainz, D. T.;
Repasky, M. P.; Knoll, E. H.; Shelley, M.; Perry, J. K.; Shaw, D. E.; Francis, P.;
Shenkin, P. S. J. Med. Chem. 2004, 47, 1739.
References and notes
18. Modeling studies were done using Sybyl 7.3–8.0 (Tripos). Energy calculations
were based on the MMFF94s force field (Halgren, T. J. Comp. Chem. 1999, 20,
720.). Protein-ligand complexes were minimized using the quasi-Newton–
Raphson procedure. For manual docking, ligands were placed in the binding
site and optimized treating all side chains within 4 Å flexible. Automated
docking was carried out using QXP (16) or Glide (17).
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19. Tautomer ratios were computed using the Tauthor approach, as implemented
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crystallized in hanging drops from conditions described by Lim et al. J. Mol. Biol.
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concentrations plus the substrate Dabcyl-c-Abu-Ile-His-Pro-Phe-His-Leu-Val-
Ile-His-Thr-EDANS at 25 M for 30 min at 37 °C and 30 min at RT in human or
l
mouse (C57BL6) plasma. Fluorescence is recorded at an excitation wavelength
of 330 nm and an emission wavelength of 485 nm. IC50 values are calculated
from inhibition as function of concentration from multiple measurements.; (b)
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Chem. Inf. Model 2010, 47, 2172.
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J. J. Tetrahedron Lett. 2006, 47, 7677.
Permeability studies using Caco-2/TC7 cells are performed using 20 lM
compound concentration in the apical to basolateral direction with a pH
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permeability value (Papp) is obtained from quantification [⁄10ꢀ7 cm/s].; (c)
Physico-chemical properties (pKa, log P, log D) are computed using MoKa (12).;
Ligand and lipophilic ligand efficiencies are computed according to: (d)
Hopkins, A. L.; Groom, C. R.; Alex, A. Drug Discovery Today 2004, 9, 430;
Leeson, P. D.; Springthorpe, B. Nat. Rev. Drug Disc. 2007, 6, 881.
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15. Inhibitory activity expressed as inhibition constant IC50 toward recombinant
human renin was determined using an assay in which the nonendogenous